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在酸、盐或温度诱导的应激环境下,与单克隆抗体C11E9和EM-7G1发生反应的单核细胞增生李斯特菌细胞抗原的表达。

Expression of cellular antigens of Listeria monocytogenes that react with monoclonal antibodies C11E9 and EM-7G1 under acid-, salt- or temperature-induced stress environments.

作者信息

Geng T, Kim K P, Gomez R, Sherman D M, Bashir R, Ladisch M R, Bhunia A K

机构信息

Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47906-2009, USA.

出版信息

J Appl Microbiol. 2003;95(4):762-72. doi: 10.1046/j.1365-2672.2003.02035.x.

Abstract

AIMS

To study the expression of cellular antigens of Listeria monocytogenes that react with monoclonal antibodies (MAbs) C11E9 and EM-7G1 under acid-, salt- or temperature-induced stress environments.

METHODS AND RESULTS

The reaction patterns of antibodies to L. monocytogenes held in stressful environments for a short duration (3 h) or grown for extended periods (16-72 h) were investigated. During both short or prolonged exposure to stress environments of high temperature (45 degrees C) and NaCl (>1.5%, w/v), reactions of whole cells of L. monocytogenes to antibodies were severely affected as determined by ELISA and by the reduced expression of the antibody-reactive 66 kDa antigen in the Western blot assay. Conversely, cold (4-15 degrees C) or acid (pH 2-3) stress environments had very little effect on antigen expression or antibody reaction. Additionally, heat-killed cells showed reduced reactions to these antibodies when compared with unheated cells. Artificially created stress environments in hotdog slurry also affected the antigen expression in L. monocytogenes. Immunoelectron microscopy revealed that the antibody-reactive antigens were uniformly present on the surface of the cells. Morphological characteristics following growth in stressed environments revealed that heat stress at 45 degrees C caused L. monocytogenes cells to be elongated and to form clumps; whereas, osmotic stress (5.5% NaCl, w/v) caused filamentous appearance with multiple septa along the length of the cell.

CONCLUSIONS

These results indicated that MAb C11E9 or EM-7G1 could detect L. monocytogenes from cold or acid-stress environments; however, they may show weaker reactions with heat or osmotically stressed cells or cells grown at 4 degrees C.

SIGNIFICANCE AND IMPACT OF THE STUDY

Bacteria in food are routinely subjected to various stresses, induced by cold, heat, salt or acid during processing and storage. Whether stresses would modify the expression of cellular antigens of L. monocytogenes is of a great concern for immunodetections in food products.

摘要

目的

研究在酸、盐或温度诱导的应激环境下,与单克隆抗体(MAb)C11E9和EM - 7G1发生反应的单核细胞增生李斯特菌细胞抗原的表达情况。

方法与结果

研究了在应激环境中短期(3小时)保存或长时间(16 - 72小时)生长的单核细胞增生李斯特菌抗体的反应模式。在短期或长期暴露于高温(45℃)和NaCl(>1.5%,w/v)的应激环境期间,通过酶联免疫吸附测定(ELISA)以及蛋白质印迹法中抗体反应性66 kDa抗原表达的降低来确定,单核细胞增生李斯特菌全细胞与抗体的反应受到严重影响。相反,冷(4 - 15℃)或酸(pH 2 - 3)应激环境对抗原表达或抗体反应影响很小。此外,与未加热的细胞相比,热灭活细胞对这些抗体的反应降低。热狗泥中人工营造的应激环境也影响了单核细胞增生李斯特菌中的抗原表达。免疫电子显微镜显示,抗体反应性抗原均匀地存在于细胞表面。在应激环境中生长后的形态学特征表明,45℃的热应激导致单核细胞增生李斯特菌细胞伸长并形成团块;而渗透应激(5.5% NaCl,w/v)导致细胞呈现丝状外观,沿细胞长度有多个隔膜。

结论

这些结果表明,单克隆抗体C11E9或EM - 7G1能够检测来自冷或酸应激环境的单核细胞增生李斯特菌;然而,它们与热应激或渗透应激细胞或在4℃下生长的细胞的反应可能较弱。

研究的意义与影响

食品中的细菌在加工和储存过程中经常受到冷、热、盐或酸诱导的各种应激。应激是否会改变单核细胞增生李斯特菌细胞抗原的表达对于食品中的免疫检测至关重要。

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